Lactoferrin: a multifunctional immunoregulatory protein?

Various immunoregulatory and anti-infective roles have been proposed for lactoferrin, the iron-binding protein present in external secretions and neutrophil secondary granules. A recent meeting¹ updated current knowledge of the structure and function of this unusual protein.     

Evidence for a novel biological role for the multifunctional β-1,3-glucan binding protein in shrimp

β-1,3-Glucan binding proteins (βGBPs) are soluble pattern recognition proteins/receptors that bind to β-1,3-glucans from fungi cell walls. In crustaceans, βGBPs are abundant plasmatic proteins produced by the hepatopancreas, and have been proved to play multiple biological functions. Here, we purified and characterized novel members of the βGBP family from the hemolymph of two Brazilian shrimps, Farfantepenaeus paulensis (FpβGBP) and Litopenaeus schmitti (LsβGBP). As observed for other crustacean species, FpβGBP and LsβGBP are monomeric proteins (～100kDa) able to enhance the activation of the prophenoloxidase system, a potent antimicrobial defense conserved in arthropods. More interestingly, we provided here evidence for a novel biological activity for shrimp βGBPs: the agglutination of fungal cells. Finally, we investigated the modulation of the βGBP gene in F. paulensis shrimps experimentally infected with a cognate fungal pathogen, Fusarium solani. From our expression data, βGBP gene is constitutively expressed in hepatopancreas and not modulated upon a non-lethal fungal infection. Herein, we have improved our knowledge about the βGBP family by the characterization of a novel biological role for this multifunctional protein in shrimp.     

Has prion protein important physiological function?

Despite the great effort aimed at uncovering the physiological function of cellular prion protein, its role remains unclear. The highly conserved amino acid sequence of PrP indicates its important function, but normally developing PrP knockout mice and cattle were prepared. Here we propose hypothesis that prion protein has no function or a redundant one and more importantly, that the conserved amino acid sequence of mammalian PrPs is not the result of their important function, but rather due to cytotoxicity of most mutations occurring in the PrP molecule. It is possible that the majority of mutations in PrP dramatically destabilizes the PrP(C) structure and causes a pathological change in conformation, so that natural selection favours individuals with non-mutated PrP.     

What establishes a protein as an allergen?

There is little known about the factors that determine the allergenicity of food proteins. Apparently, the ability of a food protein to induce an allergic response requires its presence in substantial amounts in the food supply, its durability during food processing, and its resistance to digestion in the gastrointestinal tract. In addition to the mode and degree of exposure, structural characteristics appear to play an important role for the capacity of a protein to modulate the immune response towards allergic reactions. Until now, however, there has been no indication for common structural characteristics of linear T cell or linear IgE (B cell) epitopes and the knowledge of structural characteristics of conformational IgE binding sites is very limited. Experimental data point only to certain surface areas of allergenic proteins which are important for IgE binding. Therefore, it is not possible to suggest any structural motif or conformational sequence pattern common to all allergenic proteins. Furthermore, glycosylation appears not to be a common critical determinant of allergenicity since food allergens comprise both glycoproteins and nonglycosylated proteins. Based on the few published three-dimensional structures of allergenic proteins including food proteins, one unifying feature of allergens appears to be their spherical shape. The three-dimensional structures of many more allergens have to be determined, however, to allow for a better understanding of the molecular basis of allergenicity. Most recently, new ideas have been introduced as to why certain biochemical or biologic functions such as enzymatic activities may predispose a protein to become an allergen. Proteolytically active allergens have been demonstrated to irritate the human mucosal surface, to enhance their own transmucosal uptake, and to augment IgE production. Therefore, the functional activity of some allergens may play a role among other factors in the process of sensitization and allergic responses.     

Ki67 protein: the immaculate deception?

This article updates our previous review of Ki67 published in Histopathology 10 years ago. In this period the numbers of papers published featuring this antibody has increased 10-fold from 338 to 3489 indicating the considerable enthusiasm with which this antibody has been studied. This review attempts to provide an update on the characterization of the Ki67 protein, its function and its use as a prognostic or diagnostic tool.     

Do bovine lymphocytes express a peculiar prion protein?

The cellular prion protein (PrPc) is a glycolipid-anchored cell surface protein that usually exhibits three glycosylation states. Its post-translationally modified isoform, PrPsc, is involved in the pathogenesis of various transmissible spongiform encephalopathies (TSEs). In bovine species, BSE infectivity appears to be restricted to the central nervous system; few or no detectable infectivity is found in lymphoid tissues in contrast to scrapie or variant CJD. Since expression of PrPc is a prerequisite for prion replication, we have investigated PrPc expression by bovine immune cells. Lymphocytes from blood and five different lymph organs were isolated from the same animal to assess intra- and interindividual variability of PrPc expression, considering six individuals. As shown by flow cytometry, this expression is absent or weak on granulocytes but is measurable on monocytes, B and T cells from blood and lymph organs. The activation of the bovine cells produces an upregulation of PrPc. The results of our in vitro study of PrPc biosynthesis are consistent with previous studies in other species. Interestingly, western blotting experiments showed only one form of the protein, the diglycosylated band. We propose that the glycosylation state could explain the lack of infectivity of the bovine immune cells.     

Is the pathogen of prion disease a microbial protein?

Though considerable circumstantial evidence suggests that the pathogen of prion disease is proteinaceous, it has not yet been conclusively identified. Epidemiological observations indicate that a microbial vector is responsible for the transmission of natural prion disease in sheep and goats and that the real causative agent may correspond to a structural protein of that microorganism. The microbial protein should resemble prion protein (PrP) and may replicate itself in the host by using mammalian DNA. A similar phenomenon was already described with a protein antigen of the ameba Naegleria gruberi (1–4). The various serotypes of the microbial protein may account for the existence of scrapie strains. It is proposed that many microbial proteins may be capable of replicating themselves in mammalian cells eliciting and sustaining thereby degenerative and/or autoimmune reactions subsequent to infections with microorganisms.     

How does HIV induce AIDS? The virus protein hypothesis

The genome, the structural and regulatory proteins of HIV-1, as well as the clinical course of the disease it induces are well studied. Despite this knowledge, it is still unknown how the virus induces AIDS. The disease is characterized by opportunistic infections based on a generalized and nonspecific immunosuppression. I present evidence indicating that the immunosuppressive domain of gp41 of HIV may be causually involved in the induction and progression of AIDS.     

Ischemic preconditioning: a potential role for protein S-thiolation?

Oxidant stress plays a crucial role in the triggering of cardioprotection involving ischemic preconditioning (IPC). We have used biotin-tagged cysteine to probe for redox-modified proteins in IPC protocols. Cysteine was biotinylated and introduced into isolated rat hearts. S-Thiolated proteins were detected and quantified using nonreducing western blots probed with streptavidin-horseradish peroxidase. Controls (15 min of aerobic perfusion plus 5 min of 0.5 mM biotin-cysteine plus 5 min of aerobic perfusion) showed low-level protein S-thiolation. Hearts preconditioned with 5 min of ischemia and reperfused for 5 min with biotin-cysteine plus 5 min of aerobic perfusion showed increased thiolation (160%) that was fully blocked by the antioxidant mercaptopropionylglycine, which is also known to block IPC. "Preconditioning" agonists (phorbol 12-myristate 13-acetate or phenylephrine) or oxidants (hydrogen peroxide or diamide) administered during aerobic preparations to biotin-cysteine-loaded hearts induced efficient protein S-thiolation. Preconditioning agonist-induced S-thiolation was significantly attenuated by diphenyleneiodonium (a flavoprotein inhibitor) or by the protein kinase C inhibitor bisindolylmaleimide I. Additional studies testing the role of a Nox2-containing NAD(P)H oxidase as the source of the oxidant stress essential to the triggering IPC showed that protein S-thiolation was the same in wild-type and Nox2 knockout mice.     

Targeting protein–protein interactions for cancer therapy

An increasing number of protein–protein interactions have been identified as potential intervention points for the development of anticancer agents. However, such systems have historically been considered high-risk targets due to the relatively large interaction surfaces involved in protein–protein binding. This characterization has to be reexamined as progress has been made recently in identifying small-molecule inhibitors of several protein–protein systems in oncology including the p53–MDM2 interaction. This review presents a survey of protein–protein interactions that have been identified as potential oncology targets and evaluates their attractiveness in terms of drug discovery. The analysis focuses primarily on the structural characteristics of the participating binding sites, particularly the dimensions of the sites. Known ligands are also examined, especially with regard to their druglikeness.     

Are anti-ribosomal P protein antibodies relevant in systemic lupus erythematosus?

Systemic lupus erythematosus (SLE) is a prototypal auto-immune disorder characterized with multiple organ involvement resulting in disability and increased mortality. Immune regulatory disturbances cumulate in activation of B cells and consequent auto-antibody production. Antigens for these auto-antibodies can be nuclear components and cytoplasmic elements. Anti-P antibodies react against acidic phosphorylated ribosomal proteins P0, P1, and P2 (with molecular mass of 38, 19, and 17 kDa, respectively) and are located on the S60 subunit of ribosomes. Ribosomal P proteins share a common 22-amino acid sequence that is present in the carboxyl-terminal. Anti-P antibodies can be detected in approx 15 to 20% of patients with lupus by several immunoassays, most frequently by enzyme-linked immunosorbent assay (ELISA) and/or Western blotting. However, no standardized assay is available. Auto-antibodies against eukaryotic P proteins appear highly specific for SLE; therefore, they can be used as diagnostic marker for the disease. Furthermore, association has been described with particular manifestations of lupus, especially with neuropsychiatric, renal, and hepatic involvements. Anti-P positivity and the titer of anti-P antibodies also fluctuate with clinical disease activity. Despite several lines of evidence, results are conflicting regarding the existence of such associations. Discrepancies can be explained by different study set-up or study population; it also can be attributed to the different sensitivity of tests used for the detection of anti-P antibody.